Anoctamin 5 regulates the cell cycle and affects prognosis in gastric cancer

Table 1 The 20 upregulated genes that displayed the greatest changes in their expression in anoctamin 5-depleted NUGC4 cells

Gene symbol	Gene name	Gene ID	Fold change
PLCXD1	Phosphatidylinositol-specific phospholipase C X domain containing 1	TC0X00006433.hg.1	31.97
NDRG4	NDRG family member 4	TC1600008034.hg.1	29.88
CYP3A5	Cytochrome P450 family 3 subfamily A member 5	TC0700011953.hg.1	24.81
PLCXD1	Phosphatidylinositol-specific phospholipase C X domain containing 1	TC0Y00006433.hg.1	23.44
REG4	Regenerating family member 4	TC0100015477.hg.1	19.70
CIDEB	Cell death-inducing DFFA-like effector b	TC1400008752.hg.1	17.86
DHRS9	Dehydrogenase/reductase 9	TC0200009905.hg.1	17.02
CIDEC	Cell death-inducing DFFA-like effector c	TC0300010217.hg.1	16.73
SPRR1A	Small proline rich protein 1A	TC0100010017.hg.1	16.66
APOD	Apolipoprotein D	TC0300013645.hg.1	16.57
SEMA7A	Semaphorin 7A	TC1500010018.hg.1	16.09
C11orf86	Chromosome 11 open reading frame 86	TC1100008109.hg.1	15.57
BNIPL	BCL2 interacting protein like	TC0100009936.hg.1	15.52
SUSD2	Sushi domain containing 2	TC2200006883.hg.1	15.25
MAPRE3	Microtubule-associated protein RP/EB family member 3	TC0200007048.hg.1	14.99
CYP1A1	Cytochrome P450 family 1 subfamily A member 1	TC1500010042.hg.1	14.69
GOLT1A	Golgi transport 1A	TC0100017022.hg.1	13.79
CNN2	Calponin 2	TC1900006507.hg.1	13.74
ANTXR2	ANTXR cell adhesion molecule 2	TC0400011144.hg.1	13.54
APOBEC1	Apolipoprotein B mRNA editing enzyme catalytic subunit 1	TC1200009789.hg.1	12.19

Table 2 The 20 downregulated genes that displayed the greatest changes in their expression in anoctamin 5-depleted NUGC4 cells

Gene symbol	Gene name	Gene ID	Fold change
CDK2	Cyclin-dependent kinase 2	TC1200007819.hg.1	-53.84
DTL	Denticleless E3 ubiquitin ligase homolog	TC0100011512.hg.1	-27.60
RABL3	RAB, member of RAS oncogene family like 3	TC0300012157.hg.1	-20.16
CKS1B	CDC28 protein kinase regulatory submit 1B	TC0100010100.hg.1	-19.34
LMNB1	Lamin B1	TC0500008544.hg.1	-19.18
IFRD2	Interferon-related developmental regulator 2	TC0300013981.hg.1	-18.61
PLK1	Polo like kinase 1	TC1600007235.hg.1	-17.89
GINS1	GINS complex subunit 1	TC2000007016.hg.1	-17.87
CDK6	Cyclin-dependent kinase 6	TC0700011785.hg.1	-17.08
XRCC2	X-ray repair cross complementing 2	TC0700013119.hg.1	-14.77
POGLUT3	Protein O-glucosyltransferase 3	TC1100012229.hg.1	-14.47
DSG2	Desmoglein 2	TC1800007014.hg.1	-14.37
NEMP1	Nuclear envelope integral membrane protein 1	TC1200010946.hg.1	-14.31
CBX5	Chromobox 5	TC1200010833.hg.1	-14.20
ANKRD52	Ankyrin repeat domain 52	TC1200010902.hg.1	-14.20
ITGB1	Integrin subunit beta 1	TC1000010265.hg.1	-14.16
H2BC14	H2B clustered histone 14	TC0600007377.hg.1	-13.25
SCAMP2	Secretory carrier membrane protein 2	TC1500010047.hg.1	-13.05
CMTM7	CKLF like MARVEL transmembrane domain containing 7	TC0300006968.hg.1	-13.02
GINS4	GINS complex subunit 4	TC0800007416.hg.1	-12.66

Table 3 Ingenuity pathway analysis of anoctamin 5-depleted NUGC4 cells

Category	Name	Molecules	P value (range)
Disease and Disorders	Cancer	3136	1.54 × 10-3–2.23 × 10-15
	Neurological Disease	76	1.27 × 10-3–2.23 × 10-15
	Organismal Injury and Abnormalities	3147	1.54 × 10-3–2.23 × 10-15
	Cardiovascular Disease	138	1.43 × 10-3–4.07 × 10-14
	Reproductive System Disease	1942	1.54 × 10-3–1.09 × 10-10
Molecular and Cellular Functions	DNA Replication, Recombination, and Repair	460	1.54 × 10-3–9.69 × 10-22
	Cell Cycle	650	1.54 × 10-3–1.98 × 10-20
	Cellular Assembly and Organization	533	1.57 × 10-3–9.22 × 10-16
	Cellular Development	691	1.59 × 10-3–2.66 × 10-14
	Cellular Growth and Proliferation	677	1.40 × 10-3–2.66 × 10-14
Physiological System Development and Function	Connective Tissue Development and Function	60	1.14 × 10-3–8.83 × 10-6
	Tissue Development	164	1.57 × 10-3–1.11 × 10-4
	Embryonic Development	80	1.21 × 10-3–2.07 × 10-4
	Hair and Skin Development and Function	55	6.37 × 10-4–2.07 × 10-4
	Organ Development	69	6.37 × 10-4–2.07 × 10-4

Table 4 Five-year overall survival rates with cut-off values for anoctamin 5 expression scores

Cut-off value	5-yr OS rate		P value
	Low group	High group
1.0	87.0% (n = 101)	83.7% (n = 94)	0.5136
1.1	87.7% (n = 115)	81.9% (n = 80)	0.2890
1.2	88.8% (n = 127)	78.6% (n = 68)	0.0759
1.3	89.6% (n = 137)	73.9% (n = 58)	0.0104a
1.4	88.1% (n = 142)	78.6% (n = 53)	0.0380a
1.5	87.4% (n = 152)	78.5% (n = 43)	0.0751

^aP < 0.05: Log-rank test.

OS: Overall survival.

Table 5 Relationships between clinicopathological factors of gastric cancer patients and anoctamin 5 expression

Variables		N	IHC score		P value
			High group ( 1.3)	Low group (< 1.3)
Total		195	58	137
Sex	Male	128	35	93	0.3262
	Female	67	23	44
Age	< 65	77	25	52	0.5246
	65	118	33	85
Tumor location	U	41	15	26	0.3367
	M, L	154	43	111
Tumor length (mm)	< 30	63	22	41	0.3158
	30	132	36	96
Histological type	tub1, tub2, pap	96	35	61	0.0596
	por, sig, muc	99	23	76
Lymphatic invasion	Negative	110	34	76	0.7529
	Positive	85	24	61
Venous invasion	Negative	131	38	93	0.7419
	Positive	64	20	44
pT	pT1-2	137	37	100	0.2312
	pT3-4	58	21	37
pN	pN0	139	40	99	0.7294
	pN1-3	56	18	38

IHC: Immunohistochemical; L: Lower; M: Middle; pap: muc: Mucinous adenocarcinoma; Papillary adenocarcinoma; pN: pathological N stage; por: Poorly differentiated adenocarcinoma; pT: pathological T stage; sig: Signet-ring cell carcinoma;tub: Tubular adenocarcinoma; U: Upper.

Table 6 Univariate and multivariate analyses of prognostic factors associated with 5-year overall survival

Variables	N	Univariate		Multivariate
		5-yr OS rate (%)	P value	HR (95%CI)	P value
Total	195
Sex
Male	128	87.9	0.3274
Female	67	85.3
Age
< 65	77	89.7	0.4113
65	118	85.2
Location
U	41	80.9	0.2629
M, L	154	88.6
Tumor length (mm)
< 30	63	97.8	0.0020a	3.010 (0.347-26.12)	0.3176
30	132	80.1
Histological type
tub1, tub2, pap	96	88.3	0.4544
por, sig, muc	99	85.6
Lymphatic invasion
Negative	110	96.1	0.0002a	1.872 (0.549-6.384)	0.3166
Positive	85	75.7
Venous invasion
Negative	131	90.6	0.0126a	1.085 (0.476-2.472)	0.8469
Positive	64	79.3
pT
pT1-2	137	95.9	< 0.0001a	5.240 (1.807-15.20)	0.0023c
pT3-4	58	65.5
pN
pN0	139	94.1	< 0.0001a	2.148 (0.695-6.643)	0.1844
pN1-3	56	67.6
IHC score
< 1.3	137	89.6	0.0104a	2.318 (1.016-5.288)	0.0457c
1.3	58	73.9

^aP < 0.05: Log-rank test.

^cP < 0.05: Cox hazard regression analysis.

CI: Confidence interval; HR: Hazard ratio; IHC: Immunohistochemical; L: Lower; M: Middle; muc: Mucinous adenocarcinoma; OS: Overall survival; pap: Papillary adenocarcinoma; pN: Pathological N stage; por: Poorly differentiated adenocarcinoma; pT: Pathological T stage; sig: Signet-ring cell carcinoma; tub: Tubular adenocarcinoma; U: Upper.